Figure 1

A combinatorial strategy for the expression of proteins as fusions with functional modules. A. Our toolkit’s design allows any ORF of interest to be easily expressed either on its own (above) or as a fusion (below) with N- and C-terminal modules picked from among a collection of parts (represented as multicoloured wheels). The fusion’s components are linked by standardized short peptide arms (black spheres and triangles) introduced as a result of the cloning procedure. The collection is constructed so that a given functional module can be attached either at the N-terminal or the C-terminal side of the protein of interest, which is helpful for finding the optimal arrangement in each case. B. Multiple fusions can potentially be generated by combining an ORF of interest with different modules from the collection. Specific fusions aimed at particular applications will be defined by the choice of modules used. For example, an ORF of interest N-terminally tagged with different fluorescent modules (green, yellow, blue) can be used to monitor the fusion protein’s intracellular localization under conditions best suited for each experiment. Furthermore, our versatile design allows a particular fusion protein arrangement to be cloned into vectors specific for expression in different model systems in the context of comparative studies.